Effects of direct-fed microbial supplement on ruminal and plasma metabolome of early-lactation dairy cows: Untargeted metabolomics approach.

We examined the effects of 2 multispecies direct-fed microbial (DFM) supplements on ruminal and plasma metabolome of early-lactation dairy cows using a high-coverage untargeted metabolomics approach. A total of 45 multiparous Holstein cows (41 ± 7 DIM) were enrolled for the 14-d pre-experimental and 91-d experimental period and were a subset from a lactation performance study, which used 114 cows. Cows were blocked using pre-experimental energy-corrected milk yield and randomly assigned within each block to 1 of 3 treatments: (1) corn silage-based diet with no DFM supplement (control; CON), (2) basal diet top-dressed with a mixture of Lactobacillus animalis and Propionibacterium freudenreichii at 3 × 109 cfu/d (PRO-A), or (3) basal diet top-dressed with a mixture of L. animalis, P. freudenreichii, Bacillus subtilis, and Bacillus licheniformis at 11.8 × 109 cfu/d (PRO-B). The basal diet was fed ad libitum daily as a TMR at 0600 and 1200 h for a duration of 91 d. Rumen fluid and blood samples were taken on d -3, 28, 49, 70, and 91 and immediately stored at -80°C. Before analysis, ruminal and plasma samples from d 28, 49, 70, and 91 were composited. An in-depth, untargeted metabolome profile of the composite rumen and plasma samples and the d -3 samples was developed by using a chemical isotope labeling/liquid chromatography-mass spectrometry (LC-MS)-based technique. Differentially abundant metabolites (taking into account fold change [FC] values and false discovery rates [FDR]) were identified with a volcano plot. In the rumen, compared with the CON diet, supplemental PRO-A increased (FC ≥1.2; FDR ≤0.05) the relative concentrations of 9 metabolites, including 2-hydroxy-2,4-pentadienoic acid, glutaric acid, quinolinic acid, and shikimic acid, and PRO-B increased relative concentrations of 16 metabolites, including 2-hydroxy-2,4-pentadienoic acid, glutaric acid, 16-hydroxypalmitic acid, and 2 propionate precursors (succinic and methylsuccinic acids). Relative to PRO-A, supplemental PRO-B increased (FC ≥1.2; FDR ≤0.05) relative rumen concentrations of 3 metabolites, 16-hydroxypalmitic acid, indole-3-carboxylic acid, and 5-aminopentanoic acid, but reduced relative rumen concentrations of 13 metabolites, including carnitine, threonic acid, and shikimic acid. Compared with the CON diet, relative concentrations of 13 plasma metabolites, including myxochelin A and glyceraldehyde, were increased (FC ≥1.2; FDR ≤0.05) by PRO-A supplementation, whereas those of 9 plasma metabolites, including 4-(2-aminophenyl)-2,4-dioxobutanoic acid, N-acetylornithine, and S-norlaudanosolin, were reduced (FC ≤0.83; FDR ≤0.05). Supplemental PRO-B increased (FC ≥1.2; FDR ≤0.05) relative concentrations of 9 plasma metabolites, including trans-o-hydroxybenzylidenepyruvic acid and 3-methylsalicylaldehyde, and reduced relative concentrations of 4 plasma metabolites, including ß-ethynylserine and kynurenine. Pathway analysis of the differentially abundant metabolites in both rumen and plasma revealed that these metabolites are involved in AA and fatty acid metabolism and have antimicrobial and immune-stimulating properties. The results of this study demonstrated that dietary supplementation with either PRO-A or PRO-B altered the plasma and ruminal metabolome. Notably, ruminal and plasma metabolites involved in the metabolism of AA and fatty acids and those with immunomodulatory properties were altered by either or both of the 2 microbial additives.

Effects of direct-fed microbial supplementation on performance and immune parameters of lactating dairy cows.

We evaluated the effects of supplementing bacterial direct-fed microbial (DFM) on performance, apparent total-tract digestibility, rumen fermentation, and immune parameters of lactating dairy cows. One hundred fourteen multiparous Holstein cows (41 ± 7 DIM) were used in a randomized complete block design with an experiment comprising 14 d of a covariate (pre-experimental sample and data collection) and 91 d of an experimental period. Cows were blocked based on energy-corrected milk (ECM) yield during the covariate period and the following treatments were randomly assigned within each block: (1) control (CON), corn silage-based total mixed ration without DFM; (2) PRO-A, basal diet top-dressed with a mixture of Lactobacillus animalis and Propionibacterium freudenreichii at 3 × 109 cfu/d; and 3) PRO-B, basal diet top-dressed with a mixture of L. animalis, P. freudenreichii, Bacillus subtilis, and Bacillus licheniformis at 11.8 × 109 cfu/d. Milk yield, dry matter intake (DMI), and body weight were measured daily, while milk samples for component analysis were taken on 2 consecutive days of each week of data collection. Feces, urine, rumen, and blood samples were taken during the covariate period, wk 4, 7, 10, and 13 for estimation of digestibility, N-partitioning, rumen fermentation, plasma nutrient status and immune parameters. Treatments had no effect on DMI and milk yield. Fat-corrected milk (3.5% FCM) and milk fat yield were improved with PRO-B, while milk fat percent and feed efficiency (ECM/DMI) tended to increase with PRO-B compared with PRO-A and CON. Crude fat digestibility was greater with PRO-B compared with CON. Feeding CON and PRO-A resulted in higher total volatile fatty acid concentration relative to PRO-B. Percentage of neutrophils tended to be reduced with PRO-A compared with CON and PRO-B. The mean fluorescence intensity (MFI) of anti-CD44 antibody on granulocytes tended to be higher in PRO-B compared with CON. The MFI of anti-CD62L antibody on CD8+ T cells was lower in PRO-A than PRO-B, with PRO-A also showing a tendency to be lower than CON. This study indicates the potential of DFM to improve fat digestibility with consequential improvement in fat corrected milk yield, feed efficiency and milk fat yield by lactating dairy cows. The study findings also indicate that dietary supplementation with DFM may augment immune parameters or activation of immune cells, including granulocytes and T cells; however, the overall effects on immune parameters are inconclusive.

Saturated fat supplemented in the form of triglycerides decreased digestibility and reduced performance of dairy cows as compared to calcium salt of fatty acids.

The two most popular rumen-protected fatty acid supplements in dairy cow rations are calcium salts of palm oil fatty acid calcium salts of palm oil fatty acid (CSFA) and prilled saturated fatty acids (SFAs). The objectives of this study were to determine the effects of supplementing SFA in the form of triglycerides (TSFA), as compared to CSFA, on yields, efficiency and diet digestibility in high-yielding dairy cows. Twenty-eight (14 cows in each group) multiparous cows were fed a basal diet supplemented (on DM basis) with either 12 g/kg TSFA (~350 g/cow per day - contained 980 g/kg fat; 882.3 g/kg SFAs) or 14 g/kg CSFA (~440 g/cow per day - contained 800 g/kg fat; 566.4 g/kg SFAs). The supplement amounts in the diet were balanced according to fat content. Rumen samples were taken for measurements of ammonia and volatile fatty acids concentrations, and fecal samples were taken for digestibility measurements. The CSFA cows produced 3% higher milk yields (47.6 v. 46.2 kg/day; P < 0.0001) and 4.7% higher 4% fat-corrected milk (FCM; 44.7 v. 42.7 kg/day; P = 0.02) than the TSFA cows. No difference in milk-fat content was observed, but milk-protein content was higher in the TSFA than CSFA cows. Yields of fat and protein were similar, but lactose yields were higher in TSFA cows. There were no differences in dry matter intake or efficiency calculations between groups. The ruminal ammonia concentrations were similar between groups, whereas acetate concentrations and acetate : propionate ratio were greater for CSFA than TSFA cows. The apparent total-tract digestibility of dry (P < 0.0007) and organic matters (P < 0.0003), fat (P < 0.0001), NDF and ADF (P = 0.02) were lower in the TSFA v. CSFA cows. In conclusion, the CSFA-supplemented cows produced 3% higher milk and 4.7% higher 4% FCM than the TSFA cows. However, TSFA supplementation did not depress milk-protein content. The apparent total-tract digestibility was lower for all dietary components in the TSFA cows, which was probably due to the effects of both degree of saturation and triglyceride form of the TSFA supplement. Considering that diets were balanced according to the fat content of the supplements, the lower yields of milk and FCM observed in the TSFA than CSFA cows were likely due to the lower digestibility of the fat and other nutrients in the TSFA cows, which might have negatively influenced the dietary energy content.

Symposium review: Technologies for improving fiber utilization.

The forage lignocellulosic complex is one of the greatest limitations to utilization of the nutrients and energy in fiber. Consequently, several technologies have been developed to increase forage fiber utilization by dairy cows. Physical or mechanical processing techniques reduce forage particle size and gut fill and thereby increase intake. Such techniques increase the surface area for microbial colonization and may increase fiber utilization. Genetic technologies such as brown midrib mutants (BMR) with less lignin have been among the most repeatable and practical strategies to increase fiber utilization. Newer BMR corn hybrids are better yielding than the early hybrids and recent brachytic dwarf BMR sorghum hybrids avoid lodging problems of early hybrids. Several alkalis have been effective at increasing fiber digestibility. Among these, ammoniation has the added benefit of increasing the nitrogen concentration of the forage. However, few of these have been widely adopted due to the cost and the caustic nature of the chemicals. Urea treatment is more benign but requires sufficient urease and moisture for efficacy. Ammonia-fiber expansion technology uses high temperature, moisture, and pressure to degrade lignocellulose to a greater extent than ammoniation alone, but it occurs in reactors and is therefore not currently usable on farms. Biological technologies for increasing fiber utilization such as application of exogenous fibrolytic enzymes, live yeasts, and yeast culture have had equivocal effects on forage fiber digestion in individual studies, but recent meta-analyses indicate that their overall effects are positive. Nonhydrolytic expansin-like proteins act in synergy with fibrolytic enzymes to increase fiber digestion beyond that achieved by the enzyme alone due to their ability to expand cellulose microfibrils allowing greater enzyme penetration of the cell wall matrix. White-rot fungi are perhaps the biological agents with the greatest potential for lignocellulose deconstruction, but they require aerobic conditions and several strains degrade easily digestible carbohydrates. Less ruminant nutrition research has been conducted on brown rot fungi that deconstruct lignocellulose by generating highly destructive hydroxyl radicals via the Fenton reaction. More research is needed to increase the repeatability, efficacy, cost effectiveness, and on-farm applicability of technologies for increasing fiber utilization.

Pandemic Influenza A (H1N1): knowledge among senior health workers at a secondary health care institution in Southwest, Nigeria.

OBJECTIVE: This study was conducted to assess the level of knowledge of influenza A (H1N1) infection among health care workers in a secondary health care facility in Osogbo, Southwest Nigeria. METHODS: A structured questionnaire assessing participants'knowledge of swine influenza viruses, mode of transmission, clinical criteria, primary prevention, indications for emergency care, care of affected persons and ongoing pandemic of Influenza A H1N1 was hand-delivered to all senior health care workers working in the hospital. RESULTS: The mean age of the respondents was 40.4±9.6 years. The majority (59.3%) were females. Television (73.6%) radio (61.5%), newspapers (44%), other health workers (31.9%), and internet (15.4%) were the main sources of information about influenza A H1N1. Nearly all (92.3%) felt that their source of information about the disease was inadequate. About half (51.6%) knew the virus can be transmitted from one person to another. Majority identified correctly the symptoms of infected victims such as fever (83.5%), and runny nose (79.1%). Most (87.9%) identified hand washing with soap and water as a mode of preventing transmission. Most (83.5%) felt an infected person should be isolated while very few knew oseltamivir (13.2%) and zanamivir (17.6%) are drugs to treat. Multivariate linear regression analysis identified, male sex (p=0.029), internet as a source of information (p=0.029) and knowledge of prevention of H1N1 (p= 0.005) as factors that were significantly associated with a high knowledge score on the current pandemic. CONCLUSION: There is a need to provide comprehensive information to health workers on the current pandemic.